RNAscope in-situ Hybridization

For RNAscope in-situ hybridization, mice were transcardially perfused with PBS, followed by 4% paraformaldehyde in PBS according to the manufacturer’s standard protocols. Coronal sections (20 μm) were prepared as described above. The RNAscope multiplex fluorescent reagent kit v2 and probes were from ACDbio (Minneapolis, USA). Fluorescence in-situ hybridization was performed according to the manufacturer’s standard protocols. The probes were: RNAscope Probe-Mm-Sst (Cat No. 404631), RNAscope Probe-Mm-Prkcd (Cat No. 441791), and RNAscope Probe-Mm-Fos (Cat No. 316921).